MiR-126/ABCA1 mediates exosome induced neurorestorative effects after stroke in T2DM mice

MiR-126/ABCA1 介导 T2DM 小鼠中风后外泌体诱导的神经恢复作用

• 批准号: 9339737
• 负责人: JIELI CHEN
• 金额: $ 32.81万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9339737
• 关键词: ATP-Binding Cassette Transporters; Adult; Adverse effects; Affect; Animal Model; Apolipoprotein A-I; Apolipoprotein E; Biological; Biological Process; Blood Vessels; Brain; Cells; Cerebrovascular Disorders; Cerebrum; Cholesterol; Clinical; Cognitive; Communication; Computer software; Control Groups; Data; Diabetes Mellitus; Encapsulated; Endothelial Cells; Exhibits; Female; Gene Targeting; Generations; Genes; Impairment; Injury; Ischemic Stroke; Knock-out; Lipids; Mediating; Messenger RNA; MicroRNAs; Molecular Analysis; Morbidity - disease rate; Mus; Neuraxis; Neuroglia; Neurologic; Neurological outcome; Pathway Analysis; Pathway interactions; Patients; Pattern; Play; Population; Recovery of Function; Regulator Genes; Risk Factors; Role; Serum; Signal Pathway; Stroke; Testing; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Translations; Vascular remodeling; activator 1 protein; base; brain endothelial cell; brain tissue; clinically relevant; diabetic; disability; exosome; experience; functional outcomes; genetic regulatory protein; high risk; improved; male; nanosized; neurological recovery; neurorestoration; neurovascular; novel therapeutic intervention; novel therapeutics; outcome forecast; post stroke; protein expression; response; stroke treatment; therapeutic evaluation; therapy design; white matter; white matter damage

Diabetes mellitus (DM) leads to a 3-4 fold higher risk of experiencing ischemic stroke. Stroke in type two DM (T2DM) patients and in animal models increases vascular and white matter (WM) damage in the ischemic brain, and stroke in T2DM patients has a distinct clinical pattern and a poor prognosis compared to non-DM stroke. Exosomes (Exo), are active nano size biological lipid containers, which transport regulatory genes and proteins between cells and form a major biological communication conduit, facilitating a plethora of biological responses. The regulatory molecules contained in the exosome include microRNAs (miRs), which regulate gene translation and play primary roles in mediating a vast range of biological functions. MicroRNA-126 (miR- 126) is an angiogenic microRNA and primarily expressed in endothelial cells (EC). Specific conditional knockout of EC miR-126 (miR-126EC-/-) mice have significantly worse functional outcome after stroke as well as decreased brain miR-126 and ATP-binding cassette transporter A1 (ABCA1) expression. Exosomes derived from EC (EC-Exo) have a high level of miR-126. Based on our robust preliminary data, in this pioneering study, we propose that treatment of stroke with EC-Exo will enhance neurorestorative effects after stroke in T2DM mice, possibly, via the miR-126/ABCA1 signaling pathway. This application includes three Aims. Aim-1: To test the therapeutic effects of EC-Exo on cerebral ischemic stroke in adult male and female T2DM mice. Aim-2: To evaluate whether miR-126 mediates EC-Exo treatment induced neurorestorative effects, we will evaluate the therapeutic effects of treatment of stroke in specific conditional knockout of EC miR-126 (MiR-126EC-/-) and in non-miR-126 knockout control (miR-126fl/fl) T2DM mice with EC-Exo derived from miR-126EC-/- brain ECs (miR-126EC-/-EC-Exo) or EC-Exo derived from wild type miR-126fl/fl brain ECs (miR-126fl/fl-EC-Exo) on vascular and axonal/WM remodeling and neurological and cognitive functional outcome. Aim-3: To test whether ABCA1, an indirect target of miR-126, contributes to EC-Exo treatment induced neurorestorative effects after stroke in adult male T2DM mice, mice with specific knockout of brain ABCA1 (ABCA1-B/-B) and WT ABCA1 knockout control (ABCA1fl/fl) mice will be employed. In this application, we are the first to propose that, generation of miR-126 encapsulated in EC-Exo contributes to its robust therapeutic restorative effects and that miR-126/ABCA1 pathway mediates EC-Exo-induced neurovascular and WM remodeling, and thereby improves stroke neurological and cognitive functional recovery in T2DM mice. This proposal is highly clinically relevant and if successful, will significantly impact the treatment of diabetic stroke, and possibly all stroke patients.

糖尿病 (DM) 导致缺血性中风的风险增加 3-4 倍。二型糖尿病中风 (T2DM) 患者和动物模型会增加缺血性血管和白质 (WM) 损伤 与非糖尿病患者相比，T2DM 患者的大脑和卒中具有独特的临床模式和较差的预后 中风。外泌体（Exo）是活性纳米级生物脂质容器，可运输调控基因和 细胞之间的蛋白质并形成主要的生物通讯管道，促进大量的生物 回应。外泌体中包含的调节分子包括 microRNA (miR)，它调节 基因翻译并在介导广泛的生物功能中发挥主要作用。微小RNA-126（miR- 126) 是一种血管生成 microRNA，主要在内皮细胞 (EC) 中表达。具体条件 敲除 EC miR-126 (miR-126EC-/-) 的小鼠在中风后的功能结果也明显较差 大脑 miR-126 和 ATP 结合盒转运蛋白 A1 (ABCA1) 表达减少。外泌体 源自 EC (EC-Exo) 的 miR-126 水平很高。根据我们可靠的初步数据，在此 开创性研究，我们提出用 EC-Exo 治疗中风将增强中风后的神经恢复效果 T2DM 小鼠中风的发生可能是通过 miR-126/ABCA1 信号通路实现的。该应用程序包括三个 目标。 Aim-1：测试EC-Exo对成年男性和女性缺血性脑卒中的治疗效果 T2DM 小鼠。 Aim-2：评估 miR-126 是否介导 EC-Exo 治疗诱导的神经恢复 效果，我们将评估在特定条件敲除EC中治疗中风的治疗效果 miR-126 (MiR-126EC-/-) 和具有 EC-Exo 衍生的非 miR-126 敲除对照 (miR-126fl/fl) T2DM 小鼠 来自 miR-126EC-/- 脑 EC (miR-126EC-/-EC-Exo) 或源自野生型 miR-126fl/fl 脑 EC 的 EC-Exo (miR-126fl/fl-EC-Exo) 对血管和轴突/WM 重塑以及神经和认知功能的影响 结果。 Aim-3：测试 miR-126 的间接靶标 ABCA1 是否有助于 EC-Exo 治疗 在成年雄性 T2DM 小鼠、脑特异性基因敲除小鼠中风后诱导神经恢复作用 将使用 ABCA1 (ABCA1-B/-B) 和 WT ABCA1 敲除对照 (ABCA1fl/fl) 小鼠。在这个应用程序中， 我们是第一个提出，封装在 EC-Exo 中的 miR-126 的产生有助于其稳健性 治疗恢复效果以及 miR-126/ABCA1 通路介导 EC-Exo 诱导的神经血管 和 WM 重塑，从而改善 T2DM 中风神经和认知功能恢复 老鼠。该提议具有高度的临床相关性，如果成功，将显着影响以下疾病的治疗： 糖尿病中风，可能还有所有中风患者。